Introduction
Methods
Dissections
Fixing
Antibodies
Mounting
Microscopy
Fluorescence and brightfield microscopy
After mounting, we waited an appropriate amount of time for the slides to dry. All slides were initially looked at under a Zeiss Axiovert 200 fluorescence microscope. We viewed a total of 59 stained slides under the fluorescent microscope and chose selected slides to be imaged in more detail using an Olympus Fluoview 1000 confocal microscope.
Confocal microscopy
After locating the area of interest on each slide using regular fluorescence and brightfield microscopy we used the confocal imaging software. To optimize the image we adjusted the focus, offset, voltage, and gain. To generate the images below we used a Kalman filter to minimize background noise and a longer exposure time to obtain high quality images. We additionally took Z stacks of the slides in order to view a 3D perspective of the tissue.
Images
C. borealis eye (near retina) and pericardial organ labeled with DH |
Results
2009 Class Group
Jan Plan 2009 - Advanced Neurobiology BI474
Course Goals:
- To learn techniques such as microdissection, cell culture, brightfield microscopy, fluorescence microscopy, confocal microscopy, and image analysis.
- To apply these techniques to a collaborative group project.
- To gain expertise in an area of neuroscience research; to design and conduct a research project; to analyze, interpret, and present results.
- To construct and update this website as an informative and instructive tool for the course.
Crustacean X-organ cell with neurites, 400x, Phase |
Lola the Microscope, Funded by INBRE-Maine |
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Mount Desert Island Biological Laboratory: |
Jackson Laboratory: We also toured the |
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More MDIBL Pix:
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[Above] MDIBL Coop |
[Above] Christina, Jen, Ruthie, Kimberly, Ben, Sharonda, Vic, |
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Other Laboratory Activities:
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Tilden Research Page
- 1. BI474 Protocols
- 2. General Laboratory Technique Links
- 3. Microscopy Links
- 4. Neurite Growth Research Project-JanPlan 2009
- Amy's babies
- Benwants8children.com
- Christina's Toodlers
- cool youtube video about neuron degeneration and metals
- Kimberly's Offspring
- My name is Vic- I like Neurons.
- Sharonda's Chillins
- Spring 2010 - BI474 Advanced Neurobiology